Abstract
When E. coli ribosomal subunits are reacted with 2-iminothiolane and then subjected to a mild ultraviolet irradiation, an RNA-protein cross-linking reaction occurs. About 5% of the total protein in each subunit becomes cross-linked to the RNA, and a specific sub-set of proteins is involved in the reaction. In the case of the 50S subunit, the sites of cross-linking to the 23S RNA have been determined for six of these proteins: protein L4 is cross-linked within an oligonucleotide comprising positions 613-617 in the 23S sequence, L6 within positions 2473-2481, L21 within positions 540-548, L23 within positions 137-141, L27 within positions 2332-2337 and L29 within positions 99-107.
Full text
PDF

















Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Branlant C., Krol A., Sriwidada J., Ebel J. P., Sloof P., Garrett R. A partial localisation of the binding sites of the 50 S subunit proteins L1, L20 and L23 on 23 S ribosomal RNA of Escherichia coli. FEBS Lett. 1975 Apr 1;52(2):195–201. doi: 10.1016/0014-5793(75)80805-2. [DOI] [PubMed] [Google Scholar]
- Brosius J., Dull T. J., Noller H. F. Complete nucleotide sequence of a 23S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):201–204. doi: 10.1073/pnas.77.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bäumert H. G., Sköld S. E., Kurland C. G. RNA-protein neighbourhoods of the ribosome obtained by crosslinking. Eur J Biochem. 1978 Sep 1;89(2):353–359. doi: 10.1111/j.1432-1033.1978.tb12536.x. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- Lutter L. C., Zeichhardt H., Kurland C. G. Ribosomal protein neighborhoods. I. S18 and S21 as well as S5 and S8 are neighbors. Mol Gen Genet. 1972;119(4):357–366. [PubMed] [Google Scholar]
- Maly P., Rinke J., Ulmer E., Zwieb C., Brimacombe R. Precise localization of the site of cross-linking between protein L4 and 23S ribonucleic acid induced by mild ultraviolet irradiation of Escherichia coli 50S ribosomal subunits. Biochemistry. 1980 Sep 2;19(18):4179–4188. doi: 10.1021/bi00559a007. [DOI] [PubMed] [Google Scholar]
- Mets L. J., Bogorad L. Two-dimensional polyacrylamide gel electrophoresis: an improved method for ribosomal proteins. Anal Biochem. 1974 Jan;57(1):200–210. doi: 10.1016/0003-2697(74)90065-7. [DOI] [PubMed] [Google Scholar]
- Millon R., Olomucki M., Le Gall J. Y., Golinska B., Ebel J. P., Ehresmann B. Synthesis of a new reagent, ethyl 4-azidobenzoylaminoacetimidate, and its use for RNA-protein cross-linking within Escherichia coli ribosomal 30-S subunits. Eur J Biochem. 1980 Sep;110(2):485–492. doi: 10.1111/j.1432-1033.1980.tb04890.x. [DOI] [PubMed] [Google Scholar]
- Morgan J., Brimacombe R. A series of specific ribonucleoprotein fragments from the 30-S subparticle of Escherichia coli ribosomes. Eur J Biochem. 1972 Sep 25;29(3):542–552. doi: 10.1111/j.1432-1033.1972.tb02020.x. [DOI] [PubMed] [Google Scholar]
- Möller K., Brimacombe R. Specific cross-linking of proteins S7 and L4 to ribosomal RNA, by UV irradiation of Escherichia coli ribosomal subunits. Mol Gen Genet. 1975 Dec 9;141(4):343–355. doi: 10.1007/BF00331455. [DOI] [PubMed] [Google Scholar]
- Möller K., Zwieb C., Brimacombe R. Identification of the oligonucleotide and oligopeptide involved in an RNA--protein crosslink induced by ultraviolet irradiation of Escherichia coli 30 S ribosomal subunits. J Mol Biol. 1978 Dec 15;126(3):489–506. doi: 10.1016/0022-2836(78)90055-4. [DOI] [PubMed] [Google Scholar]
- Oste C., Brimacombe R. The use of sym-triazine trichloride in RNA-protein cross-linking studies with Escherichia coli ribosomal subunits. Mol Gen Genet. 1979 Jan 5;168(1):81–86. doi: 10.1007/BF00267936. [DOI] [PubMed] [Google Scholar]
- Rinke J., Meinke M., Brimacombe R., Fink G., Rommel W., Fasold H. The use of azidoarylimidoesters in RNA-protein cross-linking studies with Escherichia coli ribosomes. J Mol Biol. 1980 Mar 5;137(3):301–304. doi: 10.1016/0022-2836(80)90318-6. [DOI] [PubMed] [Google Scholar]
- Smith K. C., Aplin R. T. A mixed photoproduct of uracil and cysteine (5-S-cysteine-6-hydrouracil). A possible model for the in vivo cross-linking of deoxyribonucleic acid and protein by ultraviolet light. Biochemistry. 1966 Jun;5(6):2125–2130. doi: 10.1021/bi00870a046. [DOI] [PubMed] [Google Scholar]
- Sommer A., Traut R. R. Identification of neighboring protein pairs in the Escherichia coli 30 S ribosomal subunit by crosslinking with methyl-4-mercaptobutyrimidate. J Mol Biol. 1976 Oct 5;106(4):995–1015. doi: 10.1016/0022-2836(76)90348-x. [DOI] [PubMed] [Google Scholar]
- Spierer P., Zimmerman R. A., Mackie G. A. RNA-protein interactions in the ribosome. Binding of 50-S-subunit proteins to 5' and 3' terminal segments of the 23-S RNA. Eur J Biochem. 1975 Apr 1;52(3):459–468. doi: 10.1111/j.1432-1033.1975.tb04014.x. [DOI] [PubMed] [Google Scholar]
- Traut R. R., Bollen A., Sun T. T., Hershey J. W., Sundberg J., Pierce L. R. Methyl 4-mercaptobutyrimidate as a cleavable cross-linking reagent and its application to the Escherichia coli 30S ribosome. Biochemistry. 1973 Aug 14;12(17):3266–3273. doi: 10.1021/bi00741a019. [DOI] [PubMed] [Google Scholar]
- Ulmer E., Meinke M., Ross A., Fink G., Brimacombe R. Chemical cross-linking of protein to RNA within intact ribosomal subunits from Escherichia coli. Mol Gen Genet. 1978 Apr 6;160(2):183–193. doi: 10.1007/BF00267480. [DOI] [PubMed] [Google Scholar]
- Volckaert G., Fiers W. Micro thin-layer techniques for rapid sequence analysis of 32P-labeled RNA: double digestion and pancreatic ribonuclease analyses. Anal Biochem. 1977 Nov;83(1):228–239. doi: 10.1016/0003-2697(77)90531-0. [DOI] [PubMed] [Google Scholar]
- Yuki A., Brimacombe R. Nucleotide sequences of Escherichia coli 16-S RNA associated with ribosomal proteins S7, S9, S10, S14 and S19. Eur J Biochem. 1975 Aug 1;56(1):23–34. doi: 10.1111/j.1432-1033.1975.tb02203.x. [DOI] [PubMed] [Google Scholar]
- Zwieb C., Brimacombe R. Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem, GFR. Nucleic Acids Res. 1979;6(5):1775–1790. doi: 10.1093/nar/6.5.1775. [DOI] [PMC free article] [PubMed] [Google Scholar]